Cracking knife and cracking components

Abstract

The present invention discloses a lysis knife and a lysis assembly, wherein the lysis knife comprises a mounting post and a tool structure, wherein the mounting post is used for being rotatably mounted in a dissociation cylinder, and the tool structure comprises a first blade fixed to the outer side surface of the mounting post, and the first blade is arranged in an up-down spiral. By driving the mounting post to rotate, the first blade is driven to rotate synchronously, so that the lysis knife rotates at a high speed in the dissociation cylinder, and since the shape of the first blade is arranged in an up-down spiral, during the forward or reverse rotation of the mounting post, the tissue block to be lysed can be squeezed downward and lifted upward, and by repeating the forward and reverse rotation actions, the tissue block can be fully lysed, which is convenient for the full dissociation of the tissue block.

Description

Technical Field

[0001] The invention relates to the technical field of cell dissociation, in particular to a lysis knife and a lysis component. Background Art

[0002] Single cell sequencing technology refers to a new technology that performs high-throughput sequencing analysis of genomes, transcriptomes, and epigenomes at the level of individual cells. It can reveal the gene structure and gene expression status of individual cells, reflect the heterogeneity between cells, and play an important role in the fields of tumors, developmental biology, microbiology, neuroscience, etc., and how to obtain single cell suspensions is the focus of current technology research and development and improvement. Summary of the invention

[0003] The main purpose of the present invention is to provide a lysis knife and a lysis component, which are intended to cut tissue blocks to lyse them, thereby ensuring the dissolution and separation effect of the dissociation solution on cells.

[0004] To achieve the above object, the present invention provides a cracking knife, comprising:

[0005] A mounting column, for rotationally mounting in the dissociation cylinder; and

[0006] The tool structure comprises a first blade fixed to the outer side surface of the mounting column, wherein the first blade is spirally arranged in an up-and-down direction.

[0007] Optionally, along the radial direction of the mounting post, the first blade has a first side portion close to the mounting post and a second side portion away from the mounting post, and an extended curve of the first side portion and an extended curve of the second side portion are intersected.

[0008] Optionally, the first blade is arranged with a width gradually increasing from top to bottom; and / or,

[0009] From top to bottom, the helix angle of the first blade gradually increases.

[0010] Optionally, a plurality of the first blades are provided, and the plurality of the first blades are spaced apart along the circumferential direction.

[0011] Optionally, an arcuate groove is provided on the lower end surface of the end of the first blade close to the mounting column, and the arcuate groove is provided through both ends along the circumferential direction, and the arcuate groove is used for the guide plate in the dissociation cylinder to pass through.

[0012] Optionally, a rib plate is provided on one side of the first blade along the circumferential direction, and the rib plate and the mounting column together enclose a guide channel, the guide channel is connected to the arc-shaped groove, and the side end of the guide channel along the circumferential direction and the lower end of the guide channel are both through-set.

[0013] Optionally, a feed channel is formed on the upper end surface of the mounting column, and a plurality of feed ports connected to the feed channel are formed on the outer side surface of the lower end of the mounting column. The plurality of feed ports are spaced apart along the circumferential direction, and the interior of the feed channel is used for the passage of a liquid extraction pipeline to discharge the homogenized solution obtained by grinding from the plurality of feed ports.

[0014] Optionally, the lower end of the mounting column is tapered downward to form an inclined surface;

[0015] A plurality of the material passing ports are arranged on the inclined surface.

[0016] Optionally, an annular groove is provided on the outer side wall of the lower end of the mounting column, and the annular groove is used to be clamped with the mounting hole at the bottom of the dissociation cylinder so that the mounting column can rotate.

[0017] The present invention further provides a lysis assembly, comprising a dissociation cylinder and a lysis knife disposed in the dissociation cylinder, wherein the lysis knife comprises:

[0018] A mounting column, for rotationally mounting in the dissociation cylinder; and

[0019] The tool structure comprises a first blade fixed to the outer side surface of the mounting column, wherein the first blade is spirally arranged in an up-and-down direction.

[0020] In the technical solution of the present invention, the mounting column is driven to rotate by a driving mechanism, thereby driving the first blade to rotate synchronously, so that the lysis knife rotates at high speed in the dissociation cylinder. Since the first blade is arranged in an up and down spiral, the tissue block to be lysed can be squeezed downward and lifted upward during the forward or reverse rotation of the mounting column. By repeating the forward and reverse rotation actions, the tissue block can be fully lysed, which facilitates the full dissociation of the tissue block. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the structures shown in these drawings without paying creative work.

[0022] Figure 1 A three-dimensional schematic diagram of an embodiment (one angle) of a lysis knife provided by the present invention;

[0023] Figure 2 for Figure 1 A three-dimensional schematic diagram of an embodiment of a cracking knife (from another angle);

[0024] Figure 3 for Figure 1 A top view of the middle cleavage knife;

[0025] Figure 4 A cross-sectional schematic diagram of an embodiment of a cracking assembly provided by the present invention.

[0026] Description of Figure Numbers:

[0027] Label name Label name 1000 Cracking components 14 Annular groove 100 Splitting knife 21 First Blade 1 Mounting Column 21a First side 11 Rib board 21b Second side 11a Guide channel 211 Arc groove 12 Feed channel 200 Dissociation Cylinder 13 Feeding port

[0028] The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0029] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] It should be noted that if a directional indication is involved in the embodiment of the present invention, the directional indication is only used to explain the relative position relationship, movement status, etc. between the components in a certain specific posture. If the specific posture changes, the directional indication will also change accordingly.

[0031] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the present invention, the descriptions of "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or suggesting their relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In addition, the technical solutions between the various embodiments can be combined with each other, but they must be based on the ability of ordinary technicians in the field to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0032] Single-cell sequencing technology refers to a new technology that performs high-throughput sequencing analysis of genomes, transcriptomes, and epigenomes at the level of individual cells. It can reveal the gene structure and gene expression status of individual cells, reflect the heterogeneity between cells, and play an important role in the fields of tumors, developmental biology, microbiology, neuroscience, etc., and how to obtain single-cell suspensions is the focus of current technology research and development and improvement.

[0033] In view of this, the present invention provides a lysis knife and a lysis component, which cut the tissue block to lyse it, thereby ensuring the dissolution and separation effect of the dissociation solution on the cells. Figures 1 to 4 An embodiment of the lysing knife provided by the present invention.

[0034] Please refer to Figures 1 to 3 The lysis knife 100 includes a mounting column 1 and a tool structure, wherein the mounting column 1 is used for being rotatably mounted in the dissociation cylinder 200, and the tool structure includes a first blade 21 fixed to the outer side surface of the mounting column 1, and the first blade 21 is spirally arranged in the up and down directions.

[0035] In the technical solution of the present invention, the mounting column 1 is driven to rotate by a driving mechanism, thereby driving the first blade 21 to rotate synchronously, so that the lysis knife 100 rotates at a high speed in the dissociation cylinder 200. Since the first blade 21 is arranged in an up and down spiral, during the forward or reverse rotation of the mounting column 1, the tissue block to be lysed can be squeezed downward and lifted upward. By repeating the forward and reverse rotation actions, the tissue block can be fully lysed, which facilitates the full dissociation of the tissue block.

[0036] It should be noted that the forward rotation direction described in the present invention is clockwise rotation, and the reverse rotation direction is counterclockwise rotation. When rotating clockwise, the first blade 21 squeezes the tissue block downward, and when rotating reversely, the first blade 21 lifts the crushed tissue upward to make it fully mixed with the dissociation solution and tissue block above. It should be noted that the time of a single clockwise rotation can be the same as or different from the time of counterclockwise rotation, and there is no limitation here.

[0037] Furthermore, the first blade 21 has a certain thickness along the radial direction of the mounting post 1. In one embodiment, the first blade 21 is twisted and extended along the same spiral direction as a whole. In another embodiment, along the radial direction of the mounting post 1, the first blade 21 has a first side portion 21a close to the mounting post 1 and a second side portion 21b away from the mounting post 1, and the extension curve of the first side portion 21a and the extension curve of the second side portion 21b are arranged to intersect. At this time, the first side portion 21a and the second side portion 21b extend along two spiral directions respectively, so that the first blade 21 is twisted as a whole. Under this structure, the first blade 21 can increase the degree of crushing of the tissue block during the process of rotating and lysing the tissue block, thereby obtaining a better lysis effect.

[0038] In one embodiment, the first blade 21 is configured to gradually increase in width from top to bottom, so that the first blade 21 is configured to be narrow at the top and wide at the bottom as a whole, which can prevent tissue blocks from being lifted up and accumulated on the top of the dissociation cylinder 200 when the mounting column 1 is reversed, while increasing the degree of crushing at the bottom.

[0039] In another embodiment, the helical angle of the first blade 21 gradually increases from top to bottom. The helical angle is the angle between the tangent of the spiral curve and the plane perpendicular to the spiral axis. This arrangement makes the first blade 21 as a whole gently sloping at the top and steeply sloping at the bottom, which can prevent the tissue blocks from being lifted up and accumulated in the upper part of the dissociation cylinder 200 when the mounting column 1 is reversed, while increasing the degree of crushing at the bottom.

[0040] It should be noted that the above two related embodiments can be set at the same time or one of them can be set selectively, and the effect is better when they are set at the same time.

[0041] The present invention does not limit the number of the first blades 21. Specifically, the first blades 21 are provided in plurality, and the plurality of first blades 21 are arranged at intervals along the circumferential direction. The intervals between the plurality of first blades 21 can be consistent or different, and can be reasonably designed according to actual needs. In this embodiment, two first blades 21 are provided, and the two first blades 21 are arranged symmetrically to ensure that each first blade 21 has sufficient spiral space.

[0042] In order to facilitate the rotational guidance of the lysis knife 100 as a whole in the dissociation cylinder 200, in this embodiment, please refer to Figure 2 The lower end surface of the first blade 21 near the end of the mounting column 1 is provided with an arc-shaped groove 211, and the arc-shaped groove 211 is provided through both ends along the circumferential direction, and the arc-shaped groove 211 is used for the guide plate in the dissociation cylinder 200 to pass through. The guide plate is always kept fixed, and the arc-shaped groove 211 rotates with the rotation of the mounting column 1, so as to achieve relative rotational cooperation between the two, and prevent the mounting column 1 from deflecting due to the existence of a matching gap under high rotation. It should be noted that the outer surface of the mounting column 1 constitutes one of the inner side surfaces of the arc-shaped groove 211.

[0043] Furthermore, in order to improve the guiding matching effect, a rib plate 11 is provided on one side of the first blade 21 along the circumferential direction, and the rib plate 11 and the mounting column 1 together enclose a guide channel 11a, and the guide channel 11a is connected to the arc-shaped groove 211, and the side end of the guide channel 11a along the circumferential direction and the lower end of the guide channel 11a are both through-set. The rib plate 11 is L-shaped as a whole, so that it naturally forms a cavity section with only an upper end air outlet with the mounting column 1. After the guide channel 11a is connected to the arc-shaped groove 211, the overall channel length that matches the guide plate is extended, thereby improving the matching stability. It should be noted that the guide plate is arranged on the bottom wall surface of the dissociation cylinder 200, is fixedly arranged, and is arc-shaped as a whole, matching the arc-shaped groove 211.

[0044] After the lysis is completed, the homogenate solution needs to be discharged and further operated in other containers. At this time, if the dissociation cylinder 200 is directly poured out, the viscous homogenate solution may be hung on the surface of the first blade 21, causing a large amount of residue. For this reason, in one embodiment of the present invention, the upper end surface of the mounting column 1 is penetrated to form a feed channel 12, and the outer side surface of the lower end of the mounting column 1 is penetrated by a plurality of feed ports 13 connected to the feed channel 12, and the plurality of feed ports 13 are distributed at intervals along the circumferential direction, and the interior of the feed channel 12 is used for the liquid extraction pipeline to pass through so as to discharge the homogenate solution obtained by grinding from the plurality of feed ports 13. After the lysis is completed, the homogenized solution is between the mounting column 1 and the inner wall of the dissociation cylinder 200. At this time, a negative pressure liquid extraction pipe can be inserted into the feed channel 12. During extraction, the homogenized solution first enters the feed channel 12 through the multiple feed ports 13, and then is sucked into the liquid extraction pipe. Such a setting can always keep the dissociation cylinder 200 fixed, and the homogenized solution can be extracted quickly and conveniently to avoid residue. It should be noted that the gap between the liquid extraction pipe and the inner wall of the feed channel 12 is reasonably set, so that the liquid extraction pipe can be inserted when the lysis knife 100 rotates, saving operating steps.

[0045] Furthermore, the lower end of the mounting column 1 is tapered downward to form an inclined surface, and a plurality of the feed openings 13 are arranged on the inclined surface. Such arrangement makes the extending direction of the feed openings 13 inclined relative to the horizontal plane, which can prevent the dissociation liquid and tissue blocks in the dissociation cylinder 200 from entering the feed channel 12 through the feed openings 13 during the rotation and lysis process of the lysis knife 100, resulting in incomplete dissociation.

[0046] In order to realize the rotation installation of the mounting column 1, an annular groove 14 is provided on the outer side wall of the lower end of the mounting column 1, and the annular groove 14 is used to be clamped with the mounting hole at the bottom of the dissociation cylinder 200, so that the mounting column 1 can rotate. Such a setting is more stable, and the clamping fit can limit the relative position of the mounting column 1 and the dissociation cylinder 200. It should be noted that although the mounting column 1 and the dissociation cylinder 200 are clamped by the shaft hole fit, it is necessary to pay attention to the movable seal to avoid leakage of the dissociation liquid.

[0047] The present invention also provides a cracking assembly 1000, please refer to Figure 4 , including a dissociation cylinder 200 and the above-mentioned lysis knife 100, the mounting column 1 of the lysis knife 100 is rotatably mounted in the dissociation cylinder 200, the dissociation cylinder 200 contains a dissociation liquid mixed with tissue blocks, the lysis assembly 1000 includes all the technical features of the above-mentioned lysis knife 100, and therefore, also has the technical effects brought by all the above-mentioned technical features, which will not be described one by one here.

[0048] Specifically, the steps for preparing the homogenate solution are as follows:

[0049] Tissue preparation: Add 1× PBS solution to a six-well plate, wash the samples stored in tissue preservation solution, take an appropriate amount of the washed samples and add them to the dissociation solution, and use surgical scissors to cut the tissue into tissue blocks of appropriate size.

[0050] Tissue lysis: Place the dissociation solution with tissue in the dissociation tube 200 and act at 37°C for 15-40 minutes, wherein the lysis knife 100 in the dissociation tube 200 needs to rotate forward every 5 minutes to break the tissue, and then the lysis knife 100 rotates reversely for the same time to incubate the tissue, so that the tissue blocks are completely digested to form a homogenized solution for the best result;

[0051] Transfer of homogenate solution: The homogenate solution is subjected to the negative pressure of the liquid extraction pipeline, and then filtered through a 40-70 μm sterile cell sieve into other containers for further operation.

[0052] The above description is only a preferred embodiment of the present invention, and does not limit the patent scope of the present invention. All equivalent structural changes made by using the contents of the present invention specification and drawings under the inventive concept of the present invention, or directly / indirectly applied in other related technical fields are included in the patent protection scope of the present invention.

Claims

1. A cracking knife, characterized in that: include: A mounting column, used for rotationally mounting in the dissociation cylinder; as well as, The tool structure comprises a first blade fixed to the outer side surface of the mounting column, wherein the first blade is spirally arranged in an up-down direction; The lower end surface of the end of the first blade close to the mounting column is provided with an arc-shaped groove, and the arc-shaped groove is provided through both ends along the circumference, and the arc-shaped groove is used for the guide plate fixedly provided on the bottom wall of the dissociation cylinder to pass through; Wherein, the outer surface of the mounting column constitutes one inner side surface of the arc-shaped groove.

2. The lysis knife according to claim 1, characterized in that: Along the radial direction of the mounting post, the first blade has a first side portion close to the mounting post and a second side portion away from the mounting post, and an extended curve of the first side portion and an extended curve of the second side portion are intersected.

3. The lysis knife according to claim 1, characterized in that: The first blade is arranged with a width gradually increasing from top to bottom; and / or, From top to bottom, the helix angle of the first blade gradually increases.

4. The lysis knife according to claim 1, characterized in that: There are a plurality of first blades, and the plurality of first blades are arranged at intervals along the circumferential direction.

5. The lysis knife according to claim 1, characterized in that: A rib plate is provided on one side of the first blade along the circumferential direction, and the rib plate and the mounting column together enclose a guide channel, the guide channel is connected to the arc-shaped groove, and the side end of the guide channel along the circumferential direction and the lower end of the guide channel are both through-set.

6. The lysis knife according to claim 1, characterized in that: The upper end surface of the mounting column is penetrated to form a feed passage, and the outer side surface of the lower end of the mounting column is penetrated with a plurality of feed ports connected to the feed passage, and the plurality of feed ports are spaced apart along the circumferential direction, and the interior of the feed passage is used for the passage of a liquid extraction pipeline to discharge the homogenized solution obtained by grinding from the plurality of feed ports.

7. The lysis knife according to claim 6, characterized in that: The lower end of the mounting column is tapered downward to form an inclined surface; A plurality of the material passing ports are arranged on the inclined surface.

8. The lysis knife according to claim 1, characterized in that: An annular groove is provided on the outer side wall of the lower end of the mounting column, and the annular groove is used for clamping with the mounting hole at the bottom of the dissociation cylinder, so that the mounting column can rotate.

9. A cracking assembly, characterized in that: It comprises a dissociation cylinder and a lysis knife arranged in the dissociation cylinder, and the lysis knife is the lysis knife as described in any one of claims 1 to 8.

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